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Literature summary extracted from

  • Di Gennaro, P.; Bruzzese, N.; Anderlini, D.; Aiossa, M.; Papacchini, M.; Campanella, L.; Bestetti, G.
    Development of microbial engineered whole-cell systems for environmental benzene determination (2010), Ecotoxicol. Environ. Saf., 74, 542-549.
    View publication on PubMed

Application

EC Number Application Comment Organism
1.3.1.19 analysis Escherichia coli, which carry genes coding for benzene dioxygenase and benzene dihydrodiol dehydrogenase, can be used to monitor benzene pollution in environmental airsamples collected from an oil refinery. The procedures involving whole-cell bioassays determine the concentration of benzene through benzene dioxygenase activity, which allows for direct correlation of oxygen consumption, and through the benzene dihydrodiol dehydrogenase that causes catechol accumulation and restores NADH necessary for the activity of the first enzyme. The assay is sensitive enough to detect the benzene vapor at a concentration level of 0.01 mM in about 30 min. The assay is applicalble to on-line monitoring of benzene concentration, no particular treatment of environmental samples is required Pseudomonas putida
1.14.12.3 analysis the enzyme is used in a whole-cell assay as biosensor for benzene concentration and vapopur, overview Pseudomonas putida

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.12.3 expression in Escherichia coli strain DH5alpha and JM109, co-expression with benzene dihydrodiol dehydrogenase under the control of the Ptac promoter or without any induction. The recombinant strains expressing the BED and the BDDH enzymes transform benzene into dihydrodiol with corresponding consumption of oxygen and regenerate NADH by converting dihydrodiol to catechol Pseudomonas putida

Protein Variants

EC Number Protein Variants Comment Organism
1.14.12.3 additional information establishing of a detection system to monitor environmental benzene contamination by co-expression of benzene dioxygenase with benzene dihydrodiol dehydrogenase in Escherichia coli. The procedures involving whole-cell bioassays determine the concentration of benzene through benzene dioxygenase activity, which allows for direct correlation of oxygen consumption, and through the benzene dihydrodiol dehydrogenase that causes catechol accumulation and restores NADH necessary for the activity of the first enzyme Pseudomonas putida

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.12.3 benzene + NADH + H+ + O2 Pseudomonas putida
-
 cis-cyclohexa-3,5-diene-1,2-diol + NAD+
-
 ?
1.14.12.3 benzene + NADH + H+ + O2 Pseudomonas putida MST
-
 cis-cyclohexa-3,5-diene-1,2-diol + NAD+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.3.1.19 Pseudomonas putida
-
-
-
1.3.1.19 Pseudomonas putida MST
-
-
-
1.14.12.3 Pseudomonas putida
-
-
-
1.14.12.3 Pseudomonas putida MST
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.12.3 benzene + NADH + H+ + O2
-
 Pseudomonas putida cis-cyclohexa-3,5-diene-1,2-diol + NAD+
-
 ?
1.14.12.3 benzene + NADH + H+ + O2
-
 Pseudomonas putida MST cis-cyclohexa-3,5-diene-1,2-diol + NAD+
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.14.12.3 BED
-
 Pseudomonas putida
1.14.12.3 benzene dioxygenase
-
 Pseudomonas putida

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.12.3 30
-
 assay at Pseudomonas putida

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.12.3 7
-
 assay at Pseudomonas putida

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.12.3 NADH
-
 Pseudomonas putida